Microfabricated Neural Probe for the Rapid Detection of Multiple Neurochemicals I

描述(申请人提供)：本申请的具体目的是测试开发一种长期的、可植入的探头的可行性，用于快速测量大脑中的多种神经化学物质。目前，神经科学研究仅限于三种测量体内神经化学物质浓度的技术；微透析法获得相对较长时间(5-20分钟)的平均浓度，酶生物传感器在相对较大的空间区域(500米长的电极)上每秒检测一种神经化学物质，以及碳纤维微电极用快速扫描循环伏安法(FSCV)检测多巴胺。需要一种新的工具来快速检测多种神经化学物质的浓度，并在细胞水平上具有空间分辨率。这样的工具将允许神经科学研究人员对疾病状态和行为背后的机制提出新的问题，例如饮酒。所提出的神经探测器满足了这一需求，它以50？m的空间分辨率每4秒检测两种神经化学物质。SB MicroSystems已经开发了一种专有的MEMS工艺，用于制造可植入的多点神经探针，用于研究大鼠的大脑。我们现有的探针具有所需的特征尺寸，空间分辨率比现有的基于酶的电极提高了10倍。建议的探测器将建立在我们现有的平台上，通过使用分子对探测器位置表面进行功能化 用于检测特定的神经化学物质。多种神经化学物质的检测将通过将不同的神经化学特定检测分子构图到相邻的探针点上来实现。我们的第一阶段应用将决定这些探针商业化的可行性：1)通过调整适配子分子修饰、固定技术和电信号检测来改进功能化探针制造，以获得最佳的灵敏度和时间响应；2)开发基于检测的恒电位电路；3)开发两个适配子分子；一个对乙醇特异、灵敏且持久，另一个用于GABA。接下来，我们将4)使用新开发的适配子将探针功能化，以检测多种分析物；5)将探针植入小鼠体内收集数据。这项第一阶段可行性研究的成功将取决于通过在体内稳定2天的探针准确检测到生理上相关的乙醇和GABA浓度。在第二阶段，我们计划开发更多的适体，这些适体可以应用于我们的探针，以检测两种以上的神经递质。我们将使用稳健设计的原则将我们的原型转化为商业产品。所附的支持信表明，我们可能能够将第一阶段的成功原型出售给神经科学研究人员。好了！

DESCRIPTION (provided by applicant): The specific aim of this application is to test the feasibility of developing a long-lasting, implantable probe for rapid measurement of multiple neurochemicals in the brain. Currently, neuroscience research is limited to three techniques for measuring the concentrations of neurochemicals in vivo; microdialysis to obtain average concentrations over a relatively long time period (5-20 minutes), enzyme-based biosensors to detect a single neurochemical every second over a relatively large spatial area (500¿m length electrode), and carbon-fiber microelectrodes to detect dopamine with fast scan cyclic voltammetry (FSCV). A new tool is required for rapid detection of concentrations of multiple neurochemicals with spatial resolution on the cellular level. Such a tool would allow neuroscience researchers to ask new questions about the mechanisms behind disease states and behaviors, such as alcohol drinking. The proposed neural probe fulfills this need by detecting two neurochemicals every 4 seconds with 50 ¿m spatial resolution. SB Microsystems has already developed a proprietary MEMS process for fabricating implantable, multi-site neural probes for studying the rat brain. Our existing probes have the feature size necessary for a 10-fold spatial resolution improvement over the available enzyme-based electrodes. The proposed probe will build on our existing platform by functionalizing the probe site surfaces with molecules for the detection of specific neurochemicals. Detection of multiple neurochemicals will be achieved by patterning different neurochemical- specific detection molecules onto adjacent probe sites. Our Phase I application will determine feasibility for commercialization of these probes by; 1) improving functionalized probe fabrication by adjusting aptamer molecule modifications, immobilization technique, and electrical signal detection to achieve the best possible sensitivity and time response, 2) developing a potentiostat circuit based for detection, 3) developing two aptamer molecules; one that is specific, sensitive, and long-lasting for ethanol, and another for GABA. Next, we will 4) functionalize the probe to detect multiple analytes with the newly develop aptamers and 5) implant probes into mice for in vivo data collection. Success in this Phase I feasibility study will be determined by the accurate detection of physiologically relevant concentrations of ethanol and GABA by probes that are stable in vivo for 2 days. In Phase II, we plan to develop more aptamers that can be applied to our probes for the detection of more than 2 neurotransmitters. We will use principles of robust design to turn our prototype into a commercial product. The attached letters of support indicate that we may be able to sell a successful prototype from Phase I to neuroscience researchers. !